The effects of the cells loading, immobilized cell efficiency, cell viability and reusability on the reaction yield on the entrapped S. cerevisiae were studied following the kinetic behavior of consecutive cycles of fermentation in batch reactions systems for free and immobilized yeast. Important parameters such as ethanol yield, productivity, sugar consumption and release biomass were reported for each reaction cycle. High ethanol yield and cells viability is observed during consecutive five fermentation cycles. The results suggest that the simple procedure described could be used to prepare reusable isolated biocatalysts for bioethanol production. We believe that, monolithic macroporous hydrogel-yeast catalyst becomes a versatile, biocompatible and reusable heterogeneous catalyst, as well as enable simplified downstream processing, able to isolate valuable strains; achieving a more eco-friendly and economically convenient process, without radical modifications of equipment and devices involve in the classical operation mode.